U.S. patent application number 12/297970 was filed with the patent office on 2009-04-23 for 3- (aminomethyliden) 2-indolinone derivates and their use as cell proliferation inhibitors.
Invention is credited to Andreas Mantoulidis, Ulrike Tontsch-Grunt, Matthias Treu.
Application Number | 20090105216 12/297970 |
Document ID | / |
Family ID | 37208327 |
Filed Date | 2009-04-23 |
United States Patent
Application |
20090105216 |
Kind Code |
A1 |
Treu; Matthias ; et
al. |
April 23, 2009 |
3- (AMINOMETHYLIDEN) 2-INDOLINONE DERIVATES AND THEIR USE AS CELL
PROLIFERATION INHIBITORS
Abstract
The present invention encompasses compounds of general formula
(1) wherein R.sup.1, R.sup.2, R.sup.3 and X are defined as in claim
1, which are suitable for the treatment of diseases characterised
by excessive or abnormal cell proliferation, and the use thereof
for preparing a pharmaceutical composition having the
above-mentioned properties. ##STR00001##
Inventors: |
Treu; Matthias; (Vienna,
AT) ; Mantoulidis; Andreas; (Wien, AT) ;
Tontsch-Grunt; Ulrike; (Baden, AT) |
Correspondence
Address: |
MICHAEL P. MORRIS;BOEHRINGER INGELHEIM USA CORPORATION
900 RIDGEBURY RD, P. O. BOX 368
RIDGEFIELD
CT
06877-0368
US
|
Family ID: |
37208327 |
Appl. No.: |
12/297970 |
Filed: |
April 23, 2007 |
PCT Filed: |
April 23, 2007 |
PCT NO: |
PCT/EP07/53959 |
371 Date: |
December 23, 2008 |
Current U.S.
Class: |
514/211.15 ;
514/235.2; 514/323; 514/418; 540/544; 544/144; 546/201;
548/486 |
Current CPC
Class: |
A61P 37/00 20180101;
A61P 31/00 20180101; A61P 31/10 20180101; A61P 33/00 20180101; A61P
35/00 20180101; A61P 37/06 20180101; A61P 19/02 20180101; A61P
13/12 20180101; A61P 29/00 20180101; A61P 15/00 20180101; A61P
17/02 20180101; C07D 405/14 20130101; C07D 405/12 20130101; A61P
7/00 20180101; A61P 9/00 20180101; A61P 19/00 20180101; A61P 17/06
20180101; C07D 491/08 20130101; C07D 413/12 20130101; C07D 487/10
20130101; C07D 401/06 20130101; A61P 31/04 20180101; A61P 1/16
20180101; A61P 17/00 20180101; C07D 401/12 20130101; A61P 1/04
20180101; C07D 403/12 20130101; A61P 35/02 20180101; A61P 31/12
20180101; A61P 43/00 20180101; A61P 21/00 20180101; C07D 209/34
20130101; A61P 25/28 20180101 |
Class at
Publication: |
514/211.15 ;
548/486; 546/201; 544/144; 540/544; 514/418; 514/323;
514/235.2 |
International
Class: |
A61K 31/404 20060101
A61K031/404; C07D 209/34 20060101 C07D209/34; C07D 401/12 20060101
C07D401/12; A61K 31/454 20060101 A61K031/454; A61K 31/553 20060101
A61K031/553; A61P 29/00 20060101 A61P029/00; A61P 37/00 20060101
A61P037/00; A61P 35/00 20060101 A61P035/00; A61K 31/5377 20060101
A61K031/5377; C07D 413/12 20060101 C07D413/12; C07D 267/10 20060101
C07D267/10 |
Foreign Application Data
Date |
Code |
Application Number |
Apr 24, 2006 |
EP |
06112985.4 |
Claims
1.) Compounds of general formula (1), ##STR00218## wherein X
denotes --NR.sup.4R.sup.5 or --OR.sup.5; and R.sup.1 denotes a
group, optionally substituted by one or more R.sup.6, selected from
among C.sub.6-15aryl and 5-15 membered heteroaryl; and R.sup.2
denotes a group, optionally substituted by one or more R.sup.6,
selected from among C.sub.3-10cycloalkyl, 3-8 membered
heterocycloalkyl, C.sub.6-15aryl and 5-15 membered heteroaryl; and
R.sup.3 denotes hydrogen or a group selected from among R.sup.a,
R.sup.b and R.sup.a substituted by one or one or more identical or
different R.sup.b and/or R.sup.c; and R.sup.4 denotes hydrogen or
C.sub.1-6alkyl; and R.sup.5 denotes hydrogen or a group, optionally
substituted by one or more R.sup.a and/or R.sup.b, selected from
among C.sub.1-6alkyl, C.sub.3-10cycloalkyl,
C.sub.4-14cycloalkylalkyl, 3-8 membered heterocycloalkyl, 4-14
membered heterocycloalkylalkyl and 6-16 membered heteroarylalkyl;
or R.sup.4 and R.sup.5 together with the nitrogen atom to which
they are linked form a heterocycloalkyl or heteroaryl ring, wherein
this ring may optionally also contain one or more identical or
different additional heteroatoms selected from among nitrogen,
oxygen and sulphur, and which may optionally be substituted by one
or more identical or different suitable R.sup.e and/or R.sup.f; and
R.sup.6 denotes a group selected from among R.sup.a, R.sup.b and
R.sup.a substituted by one or one or more identical or different
R.sup.b and/or R.sup.c; and each R.sup.a is independently of one
another selected from among C.sub.1-6alkyl, C.sub.3-10cycloalkyl,
C.sub.4-16cycloalkylalkyl, C.sub.6-10aryl, C.sub.7-16arylalkyl, 2-6
membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered
heterocycloalkylalkyl, 5-12 membered heteroaryl and 6-18 membered
heteroarylalkyl; and each R.sup.b is a suitable group, each
independently selected from among .dbd.O, --OR.sup.c,
C.sub.1-3haloalkyloxy, --OCF.sub.3, .dbd.S, --SR.sup.c,
.dbd.NR.sup.c, .dbd.NOR.sup.c, --NR.sup.cR.sup.c, halogen,
--CF.sub.3, --CN, --NC, --OCN, --SCN, --NO.sub.2, --S(O)R.sup.c,
--S(O).sub.2R.sup.c, --S(O).sub.2OR.sup.c, --S(O)NR.sup.cR.sup.c,
--S(O).sub.2NR.sup.cR.sup.c, --OS(O)R.sup.c, --OS(O).sub.2R.sup.c,
--OS(O).sub.2OR.sup.c, --OS(O).sub.2NR.sup.cR.sup.c, --C(O)R.sup.c,
--C(O)OR.sup.c, --C(O)NR.sup.cR.sup.c,
--CN(R.sup.f)NR.sup.cR.sup.c, --CN(OH)R.sup.c,
--CN(OH)NR.sup.cR.sup.c, --OC(O)R.sup.c, --OC(O)OR.sup.c,
--OC(O)NR.sup.cR.sup.c, --OCN(R.sup.f)NR.sup.cR.sup.c,
--N(R.sup.f)C(O)R.sup.c, --N(R.sup.f)C(S)R.sup.c,
--N(R.sup.f)S(O).sub.2R.sup.c, --N(R.sup.f)C(O)OR.sup.c,
--N(R.sup.f)C(O)NR.sup.cR.sup.c, --[N(R.sup.f)C(O)].sub.2R.sup.c,
--N[C(O)].sub.2R.sup.c, --N[C(O)].sub.2OR.sup.c,
--[N(R.sup.f)C(O)].sub.2OR.sup.c and
--N(R.sup.f)CN(R)NR.sup.cR.sup.c; and each R.sup.c independently of
one another is hydrogen or a group optionally substituted by one or
more identical or different R.sup.d and/or R.sup.e selected from
among C.sub.1-6alkyl, C.sub.3-10cycloalkyl,
C.sub.4-11cycloalkylalkyl, C.sub.6-10aryl, C.sub.7-16arylalkyl, 2-6
membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered
heterocycloalkylalkyl, 5-12 membered heteroaryl and 6-18 membered
heteroarylalkyl; and each R.sup.d independently of one another is
hydrogen or a group optionally substituted by one or more identical
or different R.sup.e and/or R.sup.f selected from among
C.sub.1-6alkyl, C.sub.3-8cycloalkyl, C.sub.4-11cycloalkylalkyl,
C.sub.6-10aryl, C.sub.7-16arylalkyl, 2-6 membered heteroalkyl, 3-8
membered heterocycloalkyl, 4-14 membered heterocycloalkylalkyl,
5-12 membered heteroaryl and 6-18 membered heteroarylalkyl; and
each R.sup.e is a suitable group and each independently selected
from among .dbd.O, --OR.sup.f, C.sub.1-3haloalkyloxy, --OCF.sub.3,
.dbd.S, --SR.sup.f, .dbd.NR.sup.f, .dbd.NOR.sup.f,
--NR.sup.fR.sup.f, halogen, --CF.sub.3, --CN, --NC, --OCN, --SCN,
--NO.sub.2, --S(O)R.sup.f, --S(O).sub.2R.sup.f,
--S(O).sub.2OR.sup.f, --S(O)NR.sup.fR.sup.f,
--S(O).sub.2NR.sup.fR.sup.f, --OS(O)R.sup.f, --OS(O).sub.2R.sup.f,
--OS(O).sub.2OR.sup.f, --OS(O).sub.2NR.sup.fR.sup.f, --C(O)R.sup.f,
--C(O)OR.sup.f, --C(O)NR.sup.fR.sup.f,
--CN(R.sup.g)NR.sup.fR.sup.f, --CN(OH)R.sup.f,
--C(NOH)NR.sup.fR.sup.f, --OC(O)R.sup.f, --OC(O)OR.sup.f,
--OC(O)NR.sup.fR.sup.f, --OCN(R.sup.g)NR.sup.fR.sup.f,
--N(R.sup.g)C(O)R.sup.f, --N(R.sup.g)C(S)R.sup.f,
--N(R.sup.g)S(O).sub.2R.sup.f, --N(R.sup.d)C(O)OR.sup.f,
--N(R.sup.g)C(O)NR.sup.fR.sup.f, and
--N(R.sup.g)CN(R.sup.f)NR.sup.fR.sup.f, and each R.sup.f
independently of one another is hydrogen or a group optionally
substituted by one or more identical or different R.sup.g selected
from among C.sub.1-6alkyl, C.sub.3-8cycloalkyl,
C.sub.4-11cycloalkylalkyl, C.sub.6-10aryl, C.sub.7-16arylalkyl, 2-6
membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered
heterocycloalkylalkyl, 5-12 membered heteroaryl and 6-18 membered
heteroarylalkyl; and each R.sup.g independently of one another is
hydrogen, C.sub.1-6alkyl, C.sub.3-8cycloalkyl,
C.sub.4-11cycloalkylalkyl, C.sub.6-10aryl, C.sub.7-16arylalkyl, 2-6
membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered
heterocycloalkylalkyl, 5-12 membered heteroaryl and 6-18 membered
heteroarylalkyl, optionally in the form of the tautomers, the
racemates, the enantiomers, the diastereomers and the mixtures
thereof, and optionally the pharmacologically acceptable acid
addition salts thereof, with the proviso that
3-Z-[1-(4-(piperidin-1-yl-methyl)-anilino)-1-phenyl-methylene]-5-(N-buty--
N-methyl-aminosulphonyl)-2-indolinone and
3-Z-[1-(4-(dimethylaminomethyl)-anilino)-1-phenyl-methylene]-5-aminosulph-
onyl-2-indolinone are excluded.
2.) Compounds according to claim 1, wherein R.sup.1 denotes
phenyl.
3.) Compounds according to claim 1, wherein R.sup.2 denotes phenyl,
cyclohexyl or pyridyl.
4.) Compounds according to claim 1, wherein R.sup.2 denotes
unsubstituted phenyl.
5.) Compounds according to claim 1, wherein X denotes
--NR.sup.4R.sup.5.
6.) Compounds--or the pharmaceutically effective salts
thereof--according to claim 1 for use as pharmaceutical
compositions.
7.) Compounds--or the pharmaceutically effective salts
thereof--according to claim 1 for preparing a pharmaceutical
composition with an antiproliferative activity.
8.) Pharmaceutical preparations, containing as active substance one
or more compounds of general formula (1) according to claim 1 or
the physiologically acceptable salts thereof optionally in
combination with conventional excipients and/or carriers.
9.) (canceled)
10.) Pharmaceutical preparation comprising a compound of general
formula (1) according to claim 1 and at least one other cytostatic
or cytotoxic active substance, different from formula (1),
optionally in the form of the tautomers, the racemates, the
enantiomers, the diastereomers and the mixtures thereof, and
optionally the pharmacologically acceptable acid addition salts
thereof.
11.) A method for the treatment of cancer, infection, inflammation,
autoimmune disease or any combination thereof in a warm-blooded
animal which comprises administering to the animal a
therapeutically effective amount of a compound according to claim
1.
12.) A method for the prevention of cancer, infection,
inflammation, autoimmune disease or any combination thereof in a
warm-blooded animal which comprises administering to the animal a
therapeutically effective amount of a compound according to claim
1.
Description
[0001] The present invention relates to new indolinones of general
formula (1)
##STR00002##
wherein the groups R.sup.1, R.sup.2, R.sup.3 and X have the
meanings given in the claims and specification, the isomers
thereof, processes for preparing these indolinones and their use as
pharmaceutical compositions.
[0002] Indolinones are generally known as inhibitors of kinases,
particularly with an inhibiting effect on cyclin/CDK complexes.
International Patent Application WO 01/27080 describes inter alia
indolinones which carry alkoxysulphonyl or alkylaminosulphonyl
groups in the 5-position, WO 01/16130 includes indolinones which
are substituted by an alkoxy group in the 5-position, or
indolinones that carry a methylenedioxy bridge in the 5- and
6-position. WO02/36654 describes indolinones which are substituted
by a sulphonylamino group in the 5-position.
[0003] The aim of the present invention is to indicate new active
substances that can be used for the prevention and/or treatment of
diseases characterised by excessive or abnormal cell
proliferation.
DETAILED DESCRIPTION OF THE INVENTION
[0004] It has been found that, surprisingly, compounds of general
formula (1) wherein the groups R.sup.1, R.sup.2, R.sup.3 and X have
the meanings given hereinafter, act as inhibitors of specific cell
cycle kinases. Thus the compounds according to the invention may be
used for example for the treatment of diseases associated with the
activity of specific cell cycle kinases and characterised by
excessive or abnormal cell proliferation.
[0005] The present invention relates to compounds of general
formula (1)
##STR00003##
X denotes --NR.sup.4R.sup.5 or --OR.sup.5; and R.sup.1 denotes a
group, optionally substituted by one or more R.sup.6, selected from
among C.sub.6-15aryl and 5-15 membered heteroaryl; and R.sup.2
denotes a group, optionally substituted by one or more R.sup.6,
selected from among C.sub.3-10cycloalkyl, 3-8 membered
heterocycloalkyl, C.sub.6-15aryl and 5-15 membered heteroaryl; and
R.sup.3 denotes hydrogen or a group selected from among R.sup.a,
R.sup.b and R.sup.a substituted by one or one or more identical or
different R.sup.b and/or R.sup.c; and R.sup.4 denotes hydrogen or
C.sub.1-6alkyl; and R.sup.5 denotes hydrogen or a group, optionally
substituted by one or more R.sup.a and/or R.sup.b, selected from
among C.sub.1-6alkyl, C.sub.3-10cycloalkyl,
C.sub.4-14cycloalkylalkyl, 3-8 membered heterocycloalkyl, 4-14
membered heterocycloalkylalkyl and 6-16 membered heteroarylalkyl;
or R.sup.4 and R.sup.5 together with the nitrogen atom to which
they are linked form a heterocycloalkyl or heteroaryl ring, wherein
this ring may optionally also contain one or more identical or
different additional heteroatoms, selected from among nitrogen,
oxygen and sulphur, and which may optionally be substituted by one
or more identical or different suitable R.sup.e and/or R.sup.f; and
R.sup.6 denotes a group selected from among R.sup.a, R.sup.b and
R.sup.a substituted by one or one or more identical or different
R.sup.b and/or R.sup.c; and each R.sup.a is independently of one
another selected from among C.sub.1-6alkyl, C.sub.3-10cycloalkyl,
C.sub.4-16cycloalkylalkyl, C.sub.6-10aryl, C.sub.7-16arylalkyl, 2-6
membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered
heterocycloalkylalkyl, 5-12 membered heteroaryl and 6-18 membered
heteroarylalkyl; and each R.sup.b is a suitable group, each
independently selected from among .dbd.O, --OR.sup.c,
C.sub.1-3haloalkyloxy, --OCF.sub.3, .dbd.S, --SR.sup.c,
.dbd.NR.sup.c, .dbd.NOR.sup.c, --NR.sup.cR.sup.c, halogen,
--CF.sub.3, --CN, --NC, --OCN, --SCN, --NO.sub.2, --S(O)R.sup.c,
--S(O).sub.2R.sup.c, --S(O).sub.2OR.sup.c, --S(O)NR.sup.cR.sup.c,
--S(O).sub.2NR.sup.cR.sup.c, --OS(O)R.sup.c, --OS(O).sub.2R.sup.c,
--OS(O).sub.2OR.sup.c, --OS(O).sub.2NR.sup.cR.sup.c, --C(O)R.sup.c,
--C(O)OR.sup.c, --C(O)NR.sup.cR.sup.c,
--CN(R.sup.f)NR.sup.cR.sup.c, --CN(OH)R.sup.c,
--CN(OH)NR.sup.cR.sup.c, --OC(O)R.sup.c, --OC(O)OR.sup.c,
--OC(O)NR.sup.cR.sup.c, --OCN(R.sup.f)NR.sup.cR.sup.c,
--N(R.sup.f)C(O)R.sup.c, --N(R.sup.f)C(S)R.sup.c,
--N(R.sup.f)S(O).sub.2R.sup.c, --N(R.sup.f)C(O)OR.sup.c,
--N(R.sup.f)C(O)NR.sup.cR.sup.c, --[N(R.sup.f)C(O)].sub.2R.sup.c,
--N[C(O)].sub.2R.sup.c, --N[C(O)].sub.2OR.sup.c,
--[N(R.sup.f)C(O)].sub.2OR.sup.c and
--N(R.sup.f)CN(R.sup.f)NR.sup.cR.sup.c; and each R.sup.c
independently of one another is hydrogen or a group optionally
substituted by one or more identical or different R.sup.d and/or
R.sup.e selected from among C.sub.1-6alkyl, C.sub.3-10cycloalkyl,
C.sub.4-11cycloalkylalkyl, C.sub.6-10aryl, C.sub.7-16arylalkyl, 2-6
membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered
heterocycloalkylalkyl, 5-12 membered heteroaryl and 6-18 membered
heteroarylalkyl; and each R.sup.d independently of one another is
hydrogen or a group optionally substituted by one or more identical
or different R.sup.e and/or R.sup.f selected from among
C.sub.1-6alkyl, C.sub.3-8cycloalkyl, C.sub.4-11cycloalkylalkyl,
C.sub.6-10aryl, C.sub.7-16arylalkyl, 2-6 membered heteroalkyl, 3-8
membered heterocycloalkyl, 4-14 membered heterocycloalkylalkyl,
5-12 membered heteroaryl and 6-18 membered heteroarylalkyl; and
each R.sup.e is a suitable group, each independently selected from
among .dbd.O, --OR.sup.f, C.sub.1-3haloalkyloxy, --OCF.sub.3,
.dbd.S, --SR.sup.f, .dbd.NR.sup.f, .dbd.NOR.sup.f,
--NR.sup.fR.sup.f, halogen, --CF.sub.3, --CN, --NC, --OCN, --SCN,
--NO.sub.2, --S(O)R.sup.f, --S(O).sub.2R.sup.f,
--S(O).sub.2OR.sup.f, --S(O)NR.sup.fR.sup.f,
--S(O).sub.2NR.sup.fR.sup.f, --OS(O)R.sup.f, --OS(O).sub.2R.sup.f,
--OS(O).sub.2OR.sup.f, --OS(O).sub.2NR.sup.fR.sup.f, --C(O)R.sup.f,
--C(O)OR.sup.f, --C(O)NR.sup.fR.sup.f,
--CN(R.sup.g)NR.sup.fR.sup.f, --CN(OH)R.sup.f,
--C(NOH)NR.sup.fR.sup.f, --OC(O)R.sup.f, --OC(O)OR.sup.f,
--OC(O)NR.sup.fR.sup.f, --OCN(R.sup.g)NR.sup.fR.sup.f,
--N(R.sup.g)C(O)R.sup.f, --N(R.sup.g)C(S)R.sup.f,
--N(R.sup.g)S(O).sub.2R.sup.f, --N(R.sup.d)C(O)OR.sup.f,
--N(R.sup.g)C(O)NR.sup.fR.sup.f, and
--N(R.sup.g)CN(R.sup.f)NR.sup.fR.sup.f; and each R.sup.f
independently of one another is hydrogen or a group optionally
substituted by one or more identical or different R.sup.g selected
from among C.sub.1-6alkyl, C.sub.3-8cycloalkyl,
C.sub.4-11cycloalkylalkyl, C.sub.6-10aryl, C.sub.7-16arylalkyl, 2-6
membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered
heterocycloalkylalkyl, 5-12 membered heteroaryl and 6-18 membered
heteroarylalkyl; and each R.sup.g independently of one another is
hydrogen, C.sub.1-6alkyl, C.sub.3-8cycloalkyl,
C.sub.4-11cycloalkylalkyl, C.sub.6-10aryl, C.sub.7-16arylalkyl, 2-6
membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered
heterocycloalkylalkyl, 5-12 membered heteroaryl and 6-18 membered
heteroarylalkyl, optionally in the form of the tautomers, the
racemates, the enantiomers, the diastereomers and the mixtures
thereof, and optionally the pharmacologically acceptable acid
addition salts thereof, with the proviso that [0006]
3-Z-[1-(4-(piperidin-1-yl-methyl)-anilino)-1-phenyl-methylene]-5-(N-buty--
N-methyl-aminosulphonyl)-2-indolinone and [0007]
3-Z-[1-(4-(dimethylaminomethyl)-anilino)-1-phenyl-methylene]-5-aminosulph-
onyl-2-indolinone are excluded.
(A) Aspects Relating to R.sup.1
[0008] (A1) In one aspect the invention relates to compounds of
general formula (1), wherein R.sup.1 denotes a phenyl group
optionally substituted by one or more R.sup.6.
[0009] (A2) In another aspect the invention relates to compounds of
general formula (1), wherein R.sup.1 denotes a phenyl group
substituted by an R.sup.6 in the 4-position (para position).
[0010] (A3) In another aspect the invention relates to compounds of
general formula (1), wherein R.sup.1 denotes a phenyl group
substituted by an R.sup.6 in the 4-position (para position) and
R.sup.6 denotes a group of formula
--(CH.sub.2).sub.x--NR.sup.7R.sup.8, wherein R.sup.7 and R.sup.8
each independently have the meanings of R.sup.4 and R.sup.5 and x
denotes 0 or 1.
[0011] (A4) In another aspect the invention relates to compounds of
general formula (1), wherein R.sup.1 denotes a phenyl group
substituted by an R.sup.6 in the 4-position (para position) and
R.sup.6 denotes a group of formula
--(CH.sub.2).sub.x--NR.sup.7R.sup.8, wherein
R.sup.7 denotes hydrogen or C.sub.1-3 alkyl, and R.sup.8 denotes a
group, optionally substituted by one or more R.sup.a and/or
R.sup.b, selected from among C.sub.1-3 alkyl, C.sub.3-10cycloalkyl,
C.sub.3-6 cycloalkyl-methyl, 3-8 membered heterocycloalkyl, 3-8
membered heterocycloalkyl-methyl and 5 or 6 membered
heteroarylmethyl.
[0012] (A5) In another aspect the invention relates to compounds of
general formula (1), wherein R.sup.1 denotes a phenyl group
substituted by an R.sup.6 in the 4-position (para position) and
R.sup.6 denotes a group of formula
--(CH.sub.2).sub.x--NR.sup.7R.sup.8, wherein
R.sup.7 and R.sup.8 together with the nitrogen atom to which they
are linked form a 3- to 6 membered heterocycloalkyl or 5 or 6
membered heteroaryl ring, wherein this ring may optionally also
contain one or two identical or different additional heteroatoms
selected from among nitrogen, oxygen and sulphur, and which may
optionally be substituted by a group selected from among R.sup.e
and R.sup.f.
(B) Aspects Relating to R.sup.2
[0013] (B1) In another aspect the invention relates to compounds of
general formula (1), wherein R.sup.2 denotes phenyl, cyclohexyl or
pyridyl.
[0014] (B2) In another aspect the invention relates to compounds of
general formula (1), wherein R.sup.2 denotes unsubstituted
phenyl.
(C) Aspects Relating to X
[0015] (C1) In another aspect the invention relates to compounds of
general formula (1), wherein X denotes --NR.sup.4R.sup.5.
[0016] (C2) In another aspect the invention relates to compounds of
general formula (1), wherein X denotes --NR.sup.4R.sup.5,
wherein
R.sup.4 denotes hydrogen or C.sub.1-3 alkyl, and R.sup.5 denotes a
group, optionally substituted by one or more R.sup.a and/or
R.sup.b, selected from among C.sub.1-3 alkyl, C.sub.3-10cycloalkyl,
C.sub.3-6 cycloalkyl-methyl, 3-8 membered heterocycloalkyl, 3-8
membered heterocycloalkyl-methyl and 5 or 6 membered
heteroarylmethyl.
[0017] (C3) In another aspect the invention relates to compounds of
general formula (1), wherein X denotes --NR.sup.4R.sup.5,
wherein
R.sup.4 and R.sup.5 together with the nitrogen atom to which they
are linked form a 3- to 6 membered heterocycloalkyl or 5 or 6
membered heteroaryl ring, wherein this ring may optionally also
contain one or two identical or different additional heteroatoms
selected from among nitrogen, oxygen and sulphur, and which may
optionally be substituted by a group selected from among R.sup.e
and R.sup.f.
[0018] All the above-mentioned aspects (A1) to (A5) for R.sup.1,
(B1) and (B2) for R.sup.2 and (C1) to (C3) for X may be combined
with one another as desired.
[0019] The Table below shows preferred combinations of various
aspects of the compounds of formula 1 according to the
invention:
TABLE-US-00001 embodiment R.sup.1 R.sup.2 X I-1 A1 B1 C1 I-2 A2 B1
C1 I-3 A3 B1 C1 I-4 A4 B1 C1 I-5 A5 B1 C1 I-6 A1 B2 C1 I-7 A2 B2 C2
I-8 A3 B2 C3 I-9 A4 B2 C2 I-10 A5 B2 C3
[0020] In another aspect the invention relates to compounds of
general formula (1) as pharmaceutical compositions.
[0021] In another aspect the invention relates to compounds of
general formula (1) for preparing a pharmaceutical composition with
an antiproliferative activity.
[0022] In another aspect the invention relates to a pharmaceutical
preparation, containing as active substance one or more compounds
of general formula (1) or the physiologically acceptable salts
thereof optionally in combination with conventional excipients
and/or carriers.
[0023] In another aspect the invention relates to the use of
compounds of general formula (1) for preparing a pharmaceutical
composition for the treatment and/or prevention of cancer,
infections, inflammations and autoimmune diseases.
[0024] In another aspect the invention relates to a pharmaceutical
preparation comprising a compound of general formula (1) and at
least one other cytostatic or cytotoxic active substance, different
from formula (1), optionally in the form of the tautomers, the
racemates, the enantiomers, the diastereomers and the mixtures
thereof, and optionally the pharmacologically acceptable acid
addition salts thereof.
DEFINITIONS
[0025] As used herein the following definitions apply, unless
stated otherwise.
[0026] By alkyl substituents are meant in each case saturated,
unsaturated, straight-chain or branched aliphatic hydrocarbon
groups (alkyl group) and this includes both saturated alkyl groups
and unsaturated alkenyl and alkynyl groups. Alkenyl substituents
are in each case straight-chain or branched, unsaturated alkyl
groups, which have at least one double bond. By alkynyl
substituents are meant in each case straight-chain or branched,
unsaturated alkyl groups, which have at least one triple bond.
[0027] Heteroalkyl represents unbranched or branched aliphatic
hydrocarbon chains which contain 1 to 3 heteroatoms, while each of
the available carbon and heteroatoms in the heteroalkyl chain may
optionally each be substituted independently and the heteroatoms
independently of one another are selected from among O, N, P, PO,
PO.sub.2, S, SO and SO.sub.2 (e.g. dimethylaminomethyl,
dimethylaminoethyl, dimethylaminopropyl, diethylaminomethyl,
diethylaminoethyl, diethylaminopropyl, 2-diisopropylaminoethyl,
bis-2-methoxyethylamino,
[2-(dimethylamino-ethyl)-ethyl-amino]-methyl,
3-[2-(dimethylamino-ethyl)-ethyl-amino]-propyl, hydroxymethyl,
2-hydroxyethyl, 3-hydroxypropyl, methoxy, ethoxy, propoxy,
methoxymethyl, 2-methoxyethyl).
[0028] Haloalkyl refers to alkyl groups wherein one or more
hydrogen atoms are replaced by halogen atoms. Haloalkyl includes
both saturated alkyl groups and unsaturated alkenyl and alkynyl
groups, such as for example --CF.sub.3, --CHF.sub.2, --CH.sub.2F,
--CF.sub.2CF.sub.3, --CHFCF.sub.3, --CH.sub.2CF.sub.3,
--CF.sub.2CH.sub.3, --CHFCH.sub.3, --CF.sub.2CF.sub.2CF.sub.3,
--CF.sub.2CH.sub.2CH.sub.3, --CF.dbd.CF.sub.2, --CCl.dbd.CH.sub.2,
--CBr.dbd.CH.sub.2, --CI.dbd.CH.sub.2, --C.ident.C--CF.sub.3,
--CHFCH.sub.2CH.sub.3 and --CHFCH.sub.2CF.sub.3.
[0029] Halogen refers to fluorine, chlorine, bromine and/or iodine
atoms.
[0030] By cycloalkyl is meant a mono- or polycyclic ring, wherein
the ring system may be a saturated ring but also an unsaturated,
non-aromatic ring or a spiro compound, which may optionally also
contain double bonds, such as for example cyclopropyl,
cyclopropenyl, cyclobutyl, cyclobutenyl, cyclopentyl,
cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptanyl,
cycloheptenyl, norbornyl, norbornenyl, indanyl, adamantyl,
spiroheptanyl and spiro[4.2]heptanyl.
[0031] Cycloalkylalkyl includes a non-cyclic alkyl group wherein a
hydrogen atom bound to a carbon atom is replaced by a cycloalkyl
group.
[0032] Aryl relates to monocyclic or bicyclic rings with 6-12
carbon atoms such as for example phenyl and naphthyl.
[0033] Arylalkyl includes a non-cyclic alkyl group wherein a
hydrogen atom bound to a carbon atom is replaced by an aryl
group.
[0034] By heteroaryl are meant mono- or polycyclic rings which
contain, instead of one or more carbon atoms, one or more
heteroatoms, which may be identical or different, such as e.g.
nitrogen, sulphur or oxygen atoms. Examples include furyl, furyl,
thienyl, pyrrolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl,
pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxadiazolyl,
thiadiazolyl, pyridyl, pyrimidyl, pyridazinyl, pyrazinyl and
triazinyl. Examples of bicyclic heteroaryl groups are indolyl,
isoindolyl, benzofuranyl, benzothienyl, benzoxazolyl,
benzothiazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolyl,
indazolyl, isoquinolinyl, quinolinyl, quinoxalinyl, quinolinyl,
phthalazinyl, quinazolinyl and benzotriazinyl, indolizinyl,
oxazolopyridinyl, imidazopyridinyl, naphthyridinyl, indolinyl,
isochromanyl, chromanyl, tetrahydroisoquinolinyl, isoindolinyl,
isobenzotetrahydrofuranyl, isobenzotetrahydrothienyl,
isobenzothienyl, benzoxazolyl, pyridopyridinyl,
benzotetrahydrofuranyl, benzotetrahydrothienyl, purinyl,
benzodioxolyl, triazinyl, phenoxazinyl, phenothiazinyl, pteridinyl,
benzothiazolyl, imidazopyridinyl, imidazothiazolyl,
dihydrobenzisoxazinyl, benzisoxazinyl, benzoxazinyl,
dihydrobenzisothiazinyl, benzopyranyl, benzothiopyranyl,
coumarinyl, isocoumarinyl, chromonyl, chromanonyl,
pyridinyl-N-oxide, tetrahydroquinolinyl, dihydroquinolinyl,
dihydroquinolinonyl, dihydroisoquinolinonyl, dihydrocoumarinyl,
dihydroisocoumarinyl, isoindolinonyl, benzodioxanyl,
benzoxazolinonyl, pyrrolyl-N-oxide, pyrimidinyl-N-oxide,
pyridazinyl-N-oxide, pyrazinyl-N-oxide, quinolinyl-N-oxide,
indolyl-N-oxide, indolinyl-N-oxide, isoquinolyl-N-oxide,
quinazolinyl-N-oxide, quinoxalinyl-N-oxide, phthalazinyl-N-oxide,
imidazolyl-N-oxide, isoxazolyl-N-oxide, oxazolyl-N-oxide,
thiazolyl-N-oxide, indolizinyl-N-oxide, indazolyl-N-oxide,
benzothiazolyl-N-oxide, benzimidazolyl-N-oxide, pyrrolyl-N-oxide,
oxadiazolyl-N-oxide, thiadiazolyl-N-oxide, triazolyl-N-oxide,
tetrazolyl-N-oxide, benzothiopyranyl-S-oxide and
benzothiopyranyl-S,S-dioxide.
[0035] Heteroarylalkyl encompasses a non-cyclic alkyl group wherein
a hydrogen atom bound to a carbon atom is replaced by a heteroaryl
group.
[0036] Heterocycloalkyl relates to saturated or unsaturated,
non-aromatic mono-, polycyclic or bridged polycyclic rings or spiro
compounds comprising 3-12 carbon atoms, which carry heteroatoms,
such as nitrogen, oxygen or sulphur, instead of one or more carbon
atoms. Examples of such heterocyclyl groups are tetrahydrofuranyl,
pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl,
pyrazolidinyl, pyrazolinyl, piperidinyl, piperazinyl, indolinyl,
isoindolinyl, morpholinyl, thiomorpholinyl, homomorpholinyl,
homopiperidinyl, homopiperazinyl, homothiomorpholinyl,
thiomorpholinyl-S-oxide, thiomorpholinyl-S,S-dioxide,
tetrahydropyranyl, tetrahydrothienyl,
homothiomorpholinyl-S,S-dioxide, oxazolidinonyl, dihydropyrazolyl,
dihydropyrrolyl, dihydropyrazinyl, dihydropyridinyl,
dihydropyrimidinyl, dihydrofuryl, dihydropyranyl,
tetrahydrothienyl-S-oxide, tetrahydrothienyl-S,S-dioxide,
homothiomorpholinyl-S-oxide, 6-aza-bicyclo[3.2.1]octane,
2-oxa-5-azabicyclo[2.2.1]heptane, 2-aza-bicyclo[2.2.1]hept-5-ene,
8-oxa-3-aza-bicyclo[3.2.1]octane, 3,8-diaza-bicyclo[3.2.1]octane,
2,5-diaza-bicyclo[2.2.1]heptane, 3,8-diaza-bicyclo[3.2.1]octane,
3,9-diaza-bicyclo[4.2.1]nonane and
2,6-diaza-bicyclo[3.2.2]nonane.
[0037] Heterocycloalkylalkyl relates to a non-cyclic alkyl group
wherein a hydrogen atom bound to a carbon atom is replaced by a
heterocycloalkyl group.
Preparation of the Compounds According to the Invention
Method A--2-oxo-2,3-dihydro-1H-indole-5-sulphonyl Chloride
##STR00004##
[0039] 2-indolinone (5 g, 37.6 mmol) is added batchwise at
0.degree. C. to chlorosulphonic acid (13 mL), stirred for 30 min at
this temperature and then for 16 h at RT. The reaction mixture is
slowly poured onto 200 mL of ice water, the precipitate is filtered
off, digested with water until the washing water has a neutral
reaction and dried in vacuo at 45.degree. C. Yield: 7.65 g
TABLE-US-00002 # structure educt A.1 ##STR00005## ##STR00006## A.2
##STR00007## ##STR00008##
Method B--2-oxo-2,3-dihydro-1H-indole-5-sulphonic Acid
cyclopropyl-methylamide
[0040] Cyclopropylmethylamine (5 mL, 57.6 mmol) is added dropwise
within 5 min at 0.degree. C. to a mixture of
2-oxo-2,3-dihydro-1H-indole-5-sulphonyl chloride (12 g, 51.8 mmol)
and triethylamine (10.8 mL, 77.7 mmol) in anhydrous dichloromethane
(150 mL) and stirred for 1.5 h at RT. The precipitate is filtered
off, stirred in 0.1 N HCl (150 mL) for 30 min at RT, filtered off
again, digested with water and dried at 45.degree. C. in vacuo.
Yield: 13.8 g
[0041] If during the preparation of analogous compounds the product
is not precipitated from the reaction solution in the form of a
solid, this solution is diluted with dichloromethane, washed with
0.1 N HCl, water and saturated saline solution, dried on
Na.sub.2SO.sub.4, filtered and evaporated down. The crude product
may optionally be purified by chromatography.
Method C--2-oxo-2,3-dihydro-1H-indole-5-sulphonic Acid
Ethylamide
[0042] The preparation of the sulphonamides starting from amine
hydrochlorides is carried out analogously to Method B using 3
equivalents of triethylamine.
TABLE-US-00003 # structure educt B.1 ##STR00009## ##STR00010## B.2
##STR00011## ##STR00012## B.3 ##STR00013## ##STR00014## B.4
##STR00015## ##STR00016## B.5 ##STR00017## ##STR00018## B.6
##STR00019## ##STR00020## C.1 ##STR00021## ##STR00022##
Method
D--1-benzoyl-3-[1-hydroxy-1-phenylmeth-(Z)-ylidene]-2-oxo-2,3-dihyd-
ro-1H-indole-5-sulphonic Acid Cyclopropylmethylamide
[0043] Benzoic acid (3.6 g, 29.5 mmol) and TBTU (9.8 g, 30.5 mmol)
are stirred in anhydrous DMF (10 mL) for 10 min at RT, combined
with 2-oxo-2,3-dihydro-1H-indole-5-sulphonic
acid-cyclopropylmethylamide (3.88 g, 14.5 mmol) and stirred for 5
min at RT. N-Ethyldiisopropylamine (20 mL) is added and the mixture
is stirred for 16 h at 45.degree. C. The reaction mixture is poured
onto 0.1 N HCl (150 mL), the precipitated solid is filtered off,
digested with water and dried in vacuo at 45.degree. C. Yield: 6.9
g
[0044] If during the preparation of analogous compounds the product
is not precipitated from the aqueous phase in the form of a solid,
this aqueous phase is quantitatively extracted with EtOAc. The
combined organic phases are washed with 0.1 N HCl, water, dilute
NaHCO.sub.3 solution and saturated saline solution, dried on
Na.sub.2SO.sub.4, filtered and evaporated down. The crude product
may optionally be purified by chromatography.
TABLE-US-00004 # structure educt D.1 ##STR00023## ##STR00024## D.2
##STR00025## ##STR00026## D.3 ##STR00027## ##STR00028## D.4
##STR00029## ##STR00030## D.5 ##STR00031## ##STR00032## D.6
##STR00033## ##STR00034## D.7 ##STR00035## ##STR00036## D.8
##STR00037## ##STR00038## D.9 ##STR00039## ##STR00040## D.10
##STR00041## ##STR00042## D.11 ##STR00043## ##STR00044## D.12
##STR00045## ##STR00046## D.13 ##STR00047## ##STR00048##
Method
E--1-benzoyl-3-[1-hydroxy-1-phenylmeth-(Z)-ylidene]-2-oxo-2,3-dihyd-
ro-1H-indole-5-sulphonic acid-(3-dimethylaminopropyl)amide
[0045] Benzoic acid chloride (410 .mu.L, 3.53 mmol) is added to a
mixture of 2-oxo-2,3-dihydro-1H-indole-5-sulphonic
acid-(3-dimethylamino-propyl)amide (500 mg, 1.68 mmol),
triethylamine (2.43 mL) and DMAP (20 mg) in anhydrous
dichloromethane (5 mL) and the mixture is stirred for 16 h at RT.
The working up is carried out analogously to Method D.
[0046] Yield: 511 mg
Method
F--3-[1-[4-(4-methylpiperazin-1-yl)phenylamino]-1-phenylmeth-(Z)-yl-
idene]-2-oxo-2,3-dihydro-1H-indole-5-sulphonic Acid
Cyclopropylmethylamide
[0047]
1-benzoyl-3-[1-hydroxy-1-phenylmeth-(Z)-ylidene]-2-oxo-2,3-dihydro--
1H-indole-5-sulphonic acid cyclopropylmethylamide (75 mg, 0.16
mmol), 4-(4-methylpiperazino)aniline (33.2 mg, 0.17 mmol) and
trimethylsilylimidazole (47 .mu.L, 0.32 mmol) are stirred in
anhydrous THF (500 .mu.L) for 15 min at 170.degree. C. in the
microwave. The product is filtered off, digested repeatedly with
THF and dried at 45.degree. C. in vacuo.
[0048] Yield: 30 mg
[0049] If in analogous reactions on a scale greater than 100 mg the
product is not precipitated as a solid from the reaction solution,
the latter is diluted with EtOAC, washed with 0.1 N HCl, water and
saturated saline solution, dried on Na.sub.2SO.sub.4, filtered and
evaporated down. The crude product may optionally be purified by
chromatography.
Method G--tert-butyl
4-[2-oxo-3-[1-phenyl-1-(4-pyrrolidin-1-ylmethylphenylamino)-meth-(Z)-ylid-
ene]-2,3-dihydro-1H-indole-5-sulphonylamino]-piperidine-1-carboxylate
[0050] tert.-Butyl
4-[1-benzoyl-3-[1-hydroxy-1-phenylmeth-(Z)-ylidene]-2-oxo-2,3-dihydro-1H--
indole-5-sulphonylamino]-piperidine-1-carboxylate (800 mg, 0.8
mmol), 4-pyrrolidin-1-ylmethylphenylamine (280 mg, 1.59 mmol),
chlorotrimethylsilane (206 .mu.L, 1.62 mmol) and
hexamethyldisilazane (337 .mu.L, 1.59 mmol) are refluxed in
anhydrous dioxane (8 mL) for 16 h with stirring. The reaction
mixture is evaporated down and purified by column chromatography.
Yield: 240 mg
[0051] If in analogous reactions on a scale below 100 mg the
product is not precipitated as a solid from the reaction solution,
the latter is evaporated down, the residue is taken up in DMSO, DMF
or NMP and purified by preparative HPLC/MS.
[0052] If in analogous reactions on a scale greater than 100 mg the
product is not precipitated as a solid from the reaction solution,
the latter is diluted with EtOAC, washed with 0.1 N HCl, water and
saturated saline solution, dried on Na.sub.2SO.sub.4, filtered and
evaporated down. The crude product may optionally be purified by
chromatography.
[0053] According to Method F--ethanesulphonic acid
[6-ethoxy-2-oxo-3-[1-phenyl-1-(4-pyrrolidin-1-ylmethylphenylamino)meth-(Z-
)-ylidene]-2,3-dihydro-1H-indol-5-yl]amide
[0054] Ethanesulphonic acid
[6-ethoxy-3-[1-hydroxy-1-phenyl-meth-(Z)-ylidene]-2-oxo-2,3-dihydro-1H-in-
dol-5-yl]amide (100 mg, 0.2 mmol),
4-pyrrolidin-1-ylmethylphenylamine (107 mg, 0.61 mmol) and
trimethylsilylimidazole (148 .mu.L, 1.01 mmol) are stirred in
anhydrous THF (400 .mu.L) for 15 min at 170.degree. C. in the
microwave. The mixture is diluted with DMSO, DMF or NMP, purified
by preparative HPLC/MS and the fractions obtained are lyophilised.
Yield: 2.5 mg
[0055] If in analogous reactions on a scale greater than 100 mg the
product is not precipitated as a solid from the reaction solution,
the latter is diluted with EtOAC, washed with 0.1 N HCl, water and
saturated saline solution, dried on Na.sub.2SO.sub.4, filtered and
evaporated down. The crude product may optionally be purified by
chromatography.
[0056] According to Method G--2-methylpropane-1-sulphonic acid
[3-[1-(3-methoxy-4-pyrrolidin-1-ylmethyl-phenylamino)-1-phenylmeth-(Z)-yl-
idene]-2-oxo-2,3-dihydro-1H-indol-5-yl]amide
2-methylpropane-1-sulphonic acid
[3-[1-hydroxy-1-phenylmeth-(Z)-ylidene]-2-oxo-2,3-dihydro-1H-indol-5-
-yl]amide (230 mg, 0.48 mmol),
3-methoxy-4-pyrrolidin-1-ylmethyl-phenylamine (198 mg, 0.96 mmol),
chlorotrimethylsilane (122 .mu.L, 0.96 mmol) and
hexa-methyldisilazane (203 .mu.L, 0.96 mmol) are stirred in
anhydrous THF (5 mL) for 30 min at 150.degree. C. in the microwave.
The mixture is diluted with DMSO, DMF or NMP, purified by
preparative HPLC/MS and the fractions obtained are lyophilised.
[0057] Yield: 18 mg
[0058] If in analogous reactions on a scale greater than 100 mg the
product is not precipitated as a solid from the reaction solution,
the latter is diluted with EtOAC, washed with 0.1 N HCl, water and
saturated saline solution, dried on Na.sub.2SO.sub.4, filtered and
evaporated down. The crude product may optionally be purified by
chromatography.
[0059] Reaction mixtures on the gram scale are processed at reflux
temperature.
[0060] In the event that in analogous reactions the acyl group has
not been cleaved from the indolinone-nitrogen in the course of the
reaction, the saponification is carried out with ammonia solution
or aqueous sodium hydroxide solution.
EXAMPLES 1-5
TABLE-US-00005 [0061] t.sub.Ret UV.sub.max Ex. structure [min] [nm]
[M + H].sup.+ 1 ##STR00049## 1.81 368 462.3 2 ##STR00050## 2.01 376
504.2 3 ##STR00051## 2.10 371 510.2 4 ##STR00052## 2.03 374 476.2 5
##STR00053## 2.036 387 541.3
Method J--1-(4-nitrobenzyl)pyrrolidine
##STR00054##
[0063] 4-nitrobenzylbromide (25 g, 115 mmol) is added batchwise to
a solution of pyrrolidine (24 mL, 290 mmol) in anhydrous THF (50
mL) and the mixture is stirred for 16 h at RT. The reaction mixture
is evaporated down, taken up in EtOAC (300 mL), washed with
saturated NH.sub.4Cl solution, water and saturated saline solution,
dried on sodium sulphate, filtered and evaporated down. Yield:
16.96 g
TABLE-US-00006 # structure educt J.1 ##STR00055## ##STR00056## J.2
##STR00057## ##STR00058##
Method K--4-pyrrolidin-1-ylmethylphenylamine
[0064] 1-(4-nitrobenzyl)pyrrolidine (16.96 g, 82.2 mmol) in
anhydrous THF (50 mL) is combined with Raney nickel (5 g) and
hydrogenated for 21 h under a hydrogen pressure of 7.5 bar at RT.
Optionally further catalyst is metered in and the hydrogen pressure
is readjusted as it falls. The reaction mixture is filtered,
evaporated down, combined with toluene (3.times.200 mL) and
evaporated down again. Yield: 14.46 g
TABLE-US-00007 # structure educt K.1 ##STR00059## ##STR00060## K.2
##STR00061## ##STR00062##
Method S--(2-chloro-4-nitrophenyl)methanol
##STR00063##
[0066] N,N'-carbonyldiimidazole (19.91 g, 122 mmol) is added
batchwise at RT to 2-chloro-4-nitrobenzoic acid (25 g, 90% purity,
111 mmol) in anhydrous THF (420 mL) and the mixture is stirred for
1 h. At 15-20.degree. C. sodium borohydride (13.09 g, 346 mmol) in
water (85 mL) is added dropwise and the mixture is stirred for 16 h
at RT. The reaction mixture is adjusted to pH 1 with 6 N HCl and
extracted exhaustively with EtOAc. The combined organic phases are
washed with 15% potassium carbonate solution (2.times.150 mL) and
saturated saline solution (150 mL), dried on sodium sulphate,
filtered and evaporated down. Yield: 20.6 g
TABLE-US-00008 # structure educt S.1 ##STR00064## ##STR00065## S.2
##STR00066## ##STR00067##
Method T--2-chloro-1-chloromethyl-4-nitrobenzene
[0067] (2-chloro-4-nitrophenyl)methanol (19 g, 101 mmol) is stirred
in a mixture of anhydrous dichloromethane (400 mL), thionyl
chloride (15 mL) and DMF (1 mL) for 2 h at boiling temperature. The
reaction mixture is evaporated down, the residue is taken up in
EtOAc (250 mL), washed with water (5.times.150 mL) and saturated
saline solution (150 mL), dried on sodium sulphate, filtered and
evaporated down.
[0068] Yield: 20.4 g
TABLE-US-00009 # structure educt T.1 ##STR00068## ##STR00069##
[0069] 1-(2-chloro-4-nitrobenzyl)pyrrolidine is prepared according
to Method J.
TABLE-US-00010 # structure educt J.3 ##STR00070## ##STR00071## J.4
##STR00072## ##STR00073## J.5 ##STR00074## ##STR00075##
[0070] 3-chloro-4-pyrrolidin-1-ylmethylphenylamine is prepared
according to Method K.
TABLE-US-00011 # structure educt K.3 ##STR00076## ##STR00077## K.4
##STR00078## ##STR00079## K.5 ##STR00080## ##STR00081##
[0071]
3-[1-(4-aminomethylphenylamino)-1-phenylmeth-(Z)-ylidene]-2-oxo-2,3-
-dihydro-1H-indole-5-sulphonamide derivatives are prepared
according to Method G.
EXAMPLES 6-35
TABLE-US-00012 [0072] t.sub.Ret UV.sub.max # structure [min] [nm]
[M + H].sup.+ 6 ##STR00082## 1.63 377 559.2 7 ##STR00083## 1.34 378
558.3 8 ##STR00084## 1.81 377 658.3 9 ##STR00085## 0.13 379 514.2
10 ##STR00086## 1.45 378 513.3 11 ##STR00087## 1.40 378 491.2 12
##STR00088## 1.57 379 595.2 13 ##STR00089## 1.56 381 607.2 14
##STR00090## 0.12 375 475.2 15 ##STR00091## 1.41 376 559.2 16
##STR00092## 1.38 378 574.2 17 ##STR00093## 1.59 381 565.3 18
##STR00094## 1.58 377 545.3 19 ##STR00095## 1.61 376 545.3 20
##STR00096## 1.56 379 557.3 21 ##STR00097## 1.71 378 585.5 22
##STR00098## 1.54 379 545.3 23 ##STR00099## 1.62 378 571.5 24
##STR00100## 1.32 377 477.2 25 ##STR00101## 1.69 379 567.3 26
##STR00102## 0.09 379 540.3 27 ##STR00103## 1.46 381 477.5 28
##STR00104## 1.49 378 503.3 29 ##STR00105## 1.68 380 545.3 30
##STR00106## 1.58 378 517.5 31 ##STR00107## 1.62 382 543.2 32
##STR00108## 1.55 377 505.5 33 ##STR00109## 1.59 378 531.2 34
##STR00110## 1.53 375 532.3 35 ##STR00111## 1.46 378 545.3
##STR00112##
[0073]
3-[1-[4-(tert-butyldimethylsilanyloxymethyl)phenylamino]-1-phenyl-m-
eth-(Z)-ylidene]-2-oxo-2,3-dihydro-1H-indole-5-sulphonic acid
cyclopropylmethylamide is prepared according to Method G using
(4-tert-butyldimethylsilyloxymethyl)-aniline (Wendt et al., J. Med.
Chem. 2004, 47, 303-324). The reaction solution is further reacted
directly in THF.
Method
L--3-[1-(4-hydroxymethylphenylamino)-1-phenyl-meth-(Z)-ylidene]-2-o-
xo-2,3-dihydro-1H-indole-5-sulphonic Acid
Cyclopropylmethylamide
[0074]
3-[1-[4-(tert-butyldimethylsilanyloxymethyl)phenylamino]-1-phenyl-m-
eth-(Z)-ylidene]-2-oxo-2,3-dihydro-1H-indole-5-sulphonic acid
cyclopropylmethylamide (3.7 g, 6.27 mmol) in THF (15 mL, reaction
solution from the previous step) is combined with 6 N HCl (6 mL)
and stirred for 3 h at RT. The reaction mixture is diluted with
water (150 mL) and extracted exhaustively with EtOAc. The combined
organic phases are washed with 0.1 N HCl, water, saturated
potassium carbonate solution and saturated saline solution, dried
on Na.sub.2SO.sub.4, filtered and evaporated down. Yield: 2.38
g
Method
M--4-[[[1-acetyl-5-(cyclopropylmethylsulphamoyl)-2-oxo-1,2-dihydro--
indol-(3Z)-ylidene]phenylmethyl]amino]benzyl Acetate
[0075]
3-[1-(4-hydroxymethylphenylamino)-1-phenyl-meth-(Z)-ylidene]-2-oxo--
2,3-dihydro-1H-indole-5-sulphonic acid cyclopropylmethylamide (2.38
g, 5 mmol) and acetic anhydride (1.9 mL, 20.1 mmol) are stirred in
anhydrous THF (10 mL) for 20 min at 125.degree. C. in the
microwave. The reaction mixture is diluted with dichloromethane (50
mL), washed with water and saturated saline solution, dried on
Na.sub.2SO.sub.4, filtered and evaporated down.
[0076] Yield: 2.4 g
Method
N--2-oxo-3-[1-phenyl-1-(4-pyrrolidin-1-ylmethylphenyl-amino)meth-(Z-
)-ylidene]-2,3-dihydro-1H-indole-5-sulphonic Acid
Cyclopropylmethylamide
[0077]
4-[[[1-acetyl-5-(cyclopropylmethylsulphamoyl)-2-oxo-1,2-dihydro-ind-
ol-(3Z)-ylidene]phenylmethyl]amino]benzyl acetate (80 mg, 0.14
mmol) and pyrrolidine (118 .mu.L, 1.43 mmol) are stirred in
anhydrous NMP (500 .mu.L) for 20 min at 180.degree. C. in the
microwave. The crude product is purified by preparative HPLC/MS and
freeze-dried. Yield: 16 mg Thermally unstable amines are reacted at
160-170.degree. C.
EXAMPLES 36-60
TABLE-US-00013 [0078] t.sub.Ret UV.sub.max Ex. structure [min] [nm]
[M + H].sup.+ 36 ##STR00113## 1.47 378 557.3 37 ##STR00114## 1.48
377 529.3 38 ##STR00115## 1.47 378 559.2 39 ##STR00116## 1.32 379
656.2 40 ##STR00117## 1.30 380 504.2 41 ##STR00118## 1.48 379 558.5
42 ##STR00119## 1.28 380 612.5 43 ##STR00120## 1.25 375 572.3 44
##STR00121## 1.54 377 616.2 45 ##STR00122## 1.58 379 615.2 46
##STR00123## 1.44 379 573.3 47 ##STR00124## 1.42 379 545.3 48
##STR00125## 1.52 379 543.2 49 ##STR00126## 1.45 378 558.3 50
##STR00127## 1.38 378 544.2 51 ##STR00128## 1.65 380 561.2 52
##STR00129## 1.64 375 574.5 53 ##STR00130## 1.67 375 545.3 54
##STR00131## 1.79 377 587.5 55 ##STR00132## 1.69 378 574.2 56
##STR00133## 1.68 379 547.2 57 ##STR00134## 1.57 377 560.2 58
##STR00135## 1.69 377 547.2 59 ##STR00136## 1.64 378 519.2 60
##STR00137## 1.73 377 575.5
Method H--Preparation of
1-(1-methylpiperidin-4-yl)-4-(4-nitrophenyl)piperazine
##STR00138##
[0080] 4-Fluoronitrobenzene (3 g, 21.3 mmol),
1-(1-methylpiperidin-4-yl)piperazine (3.9 g, 21.2 mmol) and
triethylamine (3.30 mL, 23.7 mmol) are stirred in anhydrous
isopropanol (10 mL) for 10 min at 160.degree. C. in the microwave.
The reaction mixture is diluted with water (10 mL), the precipitate
is filtered off washed with 50% water in isopropanol and dried in
vacuo at 45.degree. C. Yield: 5.14 g
[0081] If no crystalline product is obtained the crude mixture is
evaporated down, worked up by extraction and optionally purified by
chromatography.
TABLE-US-00014 # structure educt H.1 ##STR00139## ##STR00140## H.2
##STR00141## ##STR00142## H.3 ##STR00143## ##STR00144##
Method
I--4-[4-(1-methylpiperidin-4-yl)piperazin-1-yl]phenylamine
[0082] 1-(1-methylpiperidin-4-yl)-4-(4-nitrophenyl)piperazine (5.14
g, 16.8 mmol) is dissolved in anhydrous THF (10 mL), combined with
10% palladium on activated charcoal and hydrogenated for 17 h at 3
bar hydrogen pressure at RT. Additional catalyst is optionally
metered in and the hydrogen pressure is readjusted as it falls. The
reaction mixture is filtered, evaporated down, combined with
toluene (3.times.200 mL) and evaporated down again. Yield: 4.52
g
TABLE-US-00015 # structure educt I.1 ##STR00145## ##STR00146## I.2
##STR00147## ##STR00148## I.3 ##STR00149## ##STR00150##
[0083]
3-[1-(4-aminophenylamino)-1-phenylmeth-(Z)-ylidene]-2-oxo-2,3-dihyd-
ro-1H-indole-5-sulphonamide derivatives are prepared according to
Method G.
EXAMPLES 61-85
TABLE-US-00016 [0084] t.sub.Ret UV.sub.max Ex. structure [min] [nm]
[M + H].sup.+ 61 ##STR00151## 1.49 381 558.3 62 ##STR00152## 2.22
389 515.2 63 ##STR00153## 1.45 381 530.2 64 ##STR00154## 2.03 383
549.3 65 ##STR00155## 1.42 370 545.3 66 ##STR00156## 1.79 378 529.3
67 ##STR00157## 2.11 380 565.3 68 ##STR00158## 1.91 372 489.2 69
##STR00159## 2.29 386 565.3 70 ##STR00160## 1.49 380 661.2 71
##STR00161## 1.79 373 532.3 72 ##STR00162## 1.98 378 557.3 73
##STR00163## 1.48 381 544.5 74 ##STR00164## 1.89 389 532.3 75
##STR00165## 2.33 391 529.3 76 ##STR00166## 1.96 383 531.2 77
##STR00167## 2.08 388 531.2 78 ##STR00168## 1.52 379 562.3 79
##STR00169## 0.12 388 561.3 80 ##STR00170## 1.27 381 581.3 81
##STR00171## 0.17 379 657.3 82 ##STR00172## 1.66 384 588.3 83
##STR00173## 1.54 378 657.3 84 ##STR00174## 0.16 380 645.3 85
##STR00175## 1.65 386 588.3
[0085] Derivatives with a modification to the left-hand phenyl
nucleus are prepared in a manner which is highly analogous to the
corresponding phenyl compounds.
EXAMPLES 86-94
TABLE-US-00017 [0086] t.sub.Ret UV.sub.max Ex. structure [min] [nm]
[M + H].sup.+ 86 ##STR00176## 1.33 381 545.3 87 ##STR00177## 0.65
388 532.3 88 ##STR00178## 1.82 390 532.3 89 ##STR00179## 1.31 382
545.3 90 ##STR00180## 1.27 382 530.2 91 ##STR00181## 1.90 380 566.2
92 ##STR00182## 1.55 355 536.2 93 ##STR00183## 1.55 356 550.2 94
##STR00184## 2.11 355 537.3
Method
O--2-oxo-3-[1-phenyl-1-(4-pyrrolidin-1-ylmethylphenyl-amino)meth-(Z-
)-ylidene]-2,3-dihydro-1H-indole-5-sulphonic Acid
piperidin-4-ylamide
##STR00185##
[0088] tert-Butyl
4-[2-oxo-3-[1-phenyl-1-(4-pyrrolidin-1-ylmethylphenylamino)-meth-(Z)-ylid-
ene]-2,3-dihydro-1H-indole-5-sulphonylamino]-piperidine-1-carboxylate
(240 mg, 0.36 mmol) is stirred for 2 h in 50% trifluoroacetic acid
in dichloromethane. The reaction mixture is evaporated down and the
residue is purified by column chromatography.
[0089] Yield: 203 mg
Method
P--2-oxo-3-[1-phenyl-1-(4-pyrrolidin-1-ylmethylphenyl-amino)meth-(Z-
)-ylidene]-2,3-dihydro-1H-indole-5-sulphonic
acid-(1-acetyl-piperidin-4-yl)amide
[0090]
2-Oxo-3-[1-phenyl-1-(4-pyrrolidin-1-ylmethylphenylamino)meth-(Z)-yl-
idene]-2,3-dihydro-1H-indole-5-sulphonic acid piperidin-4-ylamide
(130 mg, 0.12 mmol) is combined in anhydrous NMP with triethylamine
(48 .mu.L, 0.35 mmol) and acetyl chloride (10 .mu.L, 1.2 mmol) and
the mixture is stirred for 12 h at RT. The reaction mixture is
purified by preparative HPLC/MS and freeze-dried. Yield: 40 mg
Method
Q--2-oxo-3-[1-phenyl-1-(4-pyrrolidin-1-ylmethylphenyl-amino)meth-(Z-
)-ylidene]-2,3-dihydro-1H-indole-5-sulphonic
acid-(1-methanesulphonylpiperidin-4-yl)amide
[0091] This is prepared analogously to Method P using
methanesulphonic acid chloride. Pyridine is optionally used instead
of triethylamine as base.
EXAMPLES 95-100
TABLE-US-00018 [0092] t.sub.Ret UV.sub.max Ex. structure [min] [nm]
[M + H].sup.+ 95 ##STR00186## 1.34 378 558.3 96 ##STR00187## 1.38
376 600.5 97 ##STR00188## 1.62 379 673.3 98 ##STR00189## 1.55 378
654.3 99 ##STR00190## 1.20 379 600.2 100 ##STR00191## 0.12 375
573.3
[0093] Methyl
4-[[[5-(N-butyl-N-methylsulphamoyl)-2-oxo-1,2-dihydroindol-(3Z)-ylidene]--
phenylmethyl]amino]benzoate is prepared according to Method F.
[0094] Yield: 150 mg
Method
U--4-[[[5-(N-butyl-N-methylsulphamoyl)-2-oxo-1,2-dihydroindol-(3Z)--
ylidene]-phenylmethyl]amino]benzoic Acid
##STR00192##
[0096] 10 N NaOH (250 .mu.L) is added to a solution of methyl
4-[[[5-(N-butyl-N-methylsulphamoyl)-2-oxo-1,2-dihydroindol-(3Z)-ylidene]--
phenylmethyl]amino]benzoate (971 mg, 1.87 mmol) in methanol (5 mL)
and the mixture is stirred for 30 min at RT. It is acidified with 1
N HCl, diluted with water (10 mL) and extracted exhaustively with
EtOAc. The combined organic phases are washed with water and
saturated saline solution, dried on Na.sub.2SO.sub.4, filtered and
evaporated down. Yield: 600 mg
Method
V--2-oxo-3-[1-phenyl-1-[4-(pyrrolidin-1-carbonyl)phenylamino]-meth--
(Z)-ylidene]-2,3-dihydro-1H-indole-5-sulphonic
Acid-N-butyl-N-methylamide
[0097]
4-[[[5-(N-butyl-N-methylsulphamoyl)-2-oxo-1,2-dihydroindol-(3Z)-yli-
dene]-phenylmethyl]amino]benzoic acid (50 mg, 0.10 mmol) in
anhydrous NMP (500 .mu.L) is combined with TBTU (47.7 mg, 0.15
mmol) and the mixture is stirred for 15 min at RT. Pyrrolidine (16
.mu.L, 0.20 mmol) and N-ethyldiisopropylamine (59 .mu.L, 0.15 mmol)
are added and the mixture is stirred for 1 h at RT. The crude
product is neutralised with formic acid, purified by preparative
HPLC/MS and freeze-dried. Yield: 16 mg
EXAMPLES 101-111
TABLE-US-00019 [0098] t.sub.Ret UV.sub.max Ex. structure [min] [nm]
[M + H].sup.+ 101 ##STR00193## 2.11 382 559.5 102 ##STR00194## 2.18
380 561.5 103 ##STR00195## 1.63 382 588.5 104 ##STR00196## 2.02 381
575.5 105 ##STR00197## 1.68 380 656.5 106 ##STR00198## 1.94 382
616.5 107 ##STR00199## 1.99 383 587.3 108 ##STR00200## 2.56 380
587.5 109 ##STR00201## 1.88 382 588.3 110 ##STR00202## 2.00 386
583.3 111 ##STR00203## 2.25 381 599.5
Method W--isobutyl 2-oxo-2,3-dihydro-1H-indole-5-sulphonate
##STR00204##
[0100] Pyridine (6 mL) is added dropwise within 5 min at 0.degree.
C. to a mixture of 2-oxo-2,3-dihydro-1H-indole-5-sulphonyl chloride
(2 g, 8.63 mmol) and 2-methyl-1-propanol (6 mL) and the mixture is
stirred for 1.5 h at RT. The reaction mixture is evaporated down,
dissolved in dichloromethane, washed with 0.1 N HCl, water and
saturated saline solution, dried on Na.sub.2SO.sub.4, filtered and
evaporated down. The crude product may optionally be purified by
chromatography. Yield: 2 g
TABLE-US-00020 # structure educt W.1 ##STR00205## ##STR00206## W.2
##STR00207## ##STR00208##
[0101]
1-benzoyl-3-[1-hydroxy-1-phenylmeth-(Z)-ylidene]-2-oxo-2,3-dihydro--
1H-indole-5-sulphonic acid esters are prepared according to Methods
D and E.
TABLE-US-00021 # structure educt E.1 ##STR00209## ##STR00210## E.2
##STR00211## ##STR00212##
[0102]
2-oxo-3-[1-phenyl-1-phenylaminometh-(Z)-ylidene]-2,3-dihydro-1H-ind-
ole-5-sulphonic acid esters are prepared according to Method G.
EXAMPLES 112-116
TABLE-US-00022 [0103] t.sub.Ret UV.sub.max Ex. structure [min] [nm]
[M + H].sup.+ 112 ##STR00213## 2.32 381 534.2 113 ##STR00214## 1.69
385 533.3 114 ##STR00215## 1.68 380 547.2 115 ##STR00216## 1.79 380
580.3 116 ##STR00217## 1.65 380 506.2
ABBREVIATIONS USED
[0104] DMAP N,N-dimethylaminopyridine [0105] DMF
N,N-dimethylformamide [0106] DMSO dimethylsulphoxide [0107] EtOAc
ethyl acetate [0108] h hour [0109] HCl hydrochloric acid [0110]
HPLC high pressure liquid chromatography [0111] M molar [0112] min
minute [0113] mL millilitre [0114] MS mass spectrometry [0115] N
normal [0116] NMP N-methylpyrrolidinone [0117] NMR nuclear
resonance spectroscopy [0118] Ph Phenyl [0119] RP reversed phase
[0120] RT ambient temperature [0121] TBTU
O-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium tetrafluoroborate
[0122] tert tertiary [0123] THF tetrahydrofuran
HPLC Methods
[0124] HPLC: Agilent 1100 Series
[0125] MS: Agilent LC/MSD SL (LCMS1: 1100 series LC/MSD)
[0126] Column: Waters, Xterra MS C18, 2.5 .mu.m, 2.1.times.30 mm,
Part. No. 186000592
[0127] Solvent: A: H.sub.2O (Millipore purified purest water) with
0.1% HCOOH [0128] B: acetonitrile (HPLC grade)
[0129] Detection: MS: Positive and negative [0130] Mass range:
120-900 m/z [0131] Fragmenter: 120 [0132] Gain EMV: 1 [0133]
Threshold: 150 [0134] Stepsize: 0.25 [0135] UV: 254 nm [0136]
Bandwide: 1 (LCMS1: 2) [0137] Reference: off
[0138] Spectrum: Range: 250-400 nm [0139] Range step: 1.00 nm
[0140] Threshold: 4.00 mAU [0141] Peakwidth: <0.01 min (LCMS1:
>0.05 min) [0142] Slit: 1 nm (LCMS 1: 2 nm)
[0143] Injection: Inj. Vol.: 5 .mu.L [0144] Inj. mode: Needle
wash
[0145] Separation: Flow: 1.10 mL/min [0146] Column temp.:
40.degree. C. [0147] Gradient: 0 min 5% solvent B [0148] 0-2.5 min
5%->95% solvent B [0149] 2.50-2.80 min 95% solvent B [0150]
2.81-3.10 min 95%->5% solvent B
[0151] The following Examples describe the biological activity of
the compounds according to the invention without restricting the
invention to these Examples.
[0152] As demonstrated by DNA staining followed by FACS or
Cellomics Array Scan analysis, the inhibition of proliferation
brought about by the compounds according to the invention is
mediated above all by errors in chromosome segregation. Because of
the accumulation of faulty segregations, massive polyploidia occurs
which may finally lead to inhibition of proliferation or even
apoptosis. On the basis of their biological properties the
compounds of general formula (I) according to the invention, their
isomers and the physiologically acceptable salts and polymorphs
thereof are suitable for treating diseases characterised by
excessive or abnormal cell proliferation.
Example Aurora-B Kinase Assay
[0153] A radioactive enzyme inhibition assay was developed using
Baculovirus-expressed recombinant human Aurora B wild-type protein
equipped at the N-terminal position with a histidine6) epitope
(His-), which is obtained from infected insect cells (SF21) and
purified.
Expression and Purification
[0154] For this, 300.times.10.sup.6 SF21 cells in SF-900II insect
cell medium (Invitrogen) are incubated for example with a suitable
amount of Baculovirus solution for 1 h at 27.degree. C. (Fernbach
flask agitator, 50 rpm). Then 250 ml SF-900 II medium is added and
agitated for 3 days (100 rpm, 27.degree. C.). Three hours before
harvesting, okadaic acid (C.sub.44H.sub.68O.sub.13, Calbiochem
#495604) is added to the culture (final concentration 0.1 .mu.M) in
order to stabilise phosphorylation sites on recombinant Aurora B.
The cells are pelleted by centrifugation (1000 rpm, 5 min,
4.degree. C.), the supernatant is discarded and the pellet is
frozen in liquid nitrogen. The pellet is thawed (37.degree. C., 5
min) and resuspended in lysis buffer. 40 mL lysis buffer (25 mM
Tris/Cl, 10 mM MgCl.sub.2, 300 mM NaCl, 20 mM imidazole, pH 8.0,
0.07% 2-mercaptoethanol and Protease-Inhibitor-Complete from Roche
Diagnostics) is used for 200 mL of volume of the starting culture.
After two rapid freezing/thawing cycles (liquid nitrogen at
37.degree. C.), the lysate is kept on ice for 30 min, then
incubated (2 h, 4.degree. C.) with washed Ni-NTA beads (Ni-NTA
Superflow Beads, 4 mL per 200 mL of starting culture) and placed in
an Econo-Pac column (Biorad #732-1010). Five washes with in each
case 10 column volumes of washing buffer (25 mM Tris/Cl, 10 mM
MgCl.sub.2, 1000 mM NaCl, 20 mM imidazole, pH 8.0, 0.07%
2-mercaptoethanol and Protease-Inhibitor-Complete from Roche
Diagnostics) precede the elution in 8 ml (per 200 ml of starting
culture) elution buffer (25 mM Tris/Cl pH 8.0, 300 mM NaCl, 10 mM
MgCl.sub.2, 0.03% Brij-35, 10% glycerol, 0.07% 2-mercaptoethanol,
400 mM imidazole). The combined eluate fractions are desalinated
using a Sephadex G25 column and transferred into freezing buffer
(50 mM tris/Cl pH 8.0, 150 mM NaCl, 0.1 mM EDTA, 0.03% Brij-35, 10%
glycerol, 1 mM DTT).
Kinase Assay
[0155] Test substances are placed in a polypropylene dish (96
wells, Greiner #655 201), in order to cover a concentration frame
of 10 .mu.M-0.0001 .mu.M. The final concentration of DMSO in the
assay is 5%. 30 .mu.L of protein mix (50 mM tris/Cl pH 7.5, 25 mM
MgCl.sub.2, 25 mM NaCl, 167 .mu.M ATP, 200 ng His-Aurora B in
freezing buffer) are pipetted into the 10 .mu.l of test substance
provided in 25% DMSO and this is incubated for 15 min at RT. Then
10 .mu.L of peptide mix (100 mM tris/Cl pH 7.5, 50 mM MgCl.sub.2,
50 mM NaCl, 5 .mu.M NaF, 5 .mu.M DTT, 1 .mu.Ci gamma-P33-ATP
[Amersham], 50 .mu.M substrate peptide [biotin-EPLERRLSLVPDS or
multimers thereof, or biotin-EPLERRLSLVPKM or multimers thereof, or
biotin-LRRWSLGLRRWSLGLRRWSLGLRRWSLG]) are added. The reaction is
incubated for 75 min (ambient temperature) and stopped by the
addition of 180 .mu.L of 6.4% trichloroacetic acid and incubated
for 20 min on ice. A multiscreen filtration plate (Millipore, MAIP
NOB 10) is equilibrated first of all with 100 .mu.L 70% ethanol and
then with 180 .mu.L trichloroacetic acid and the liquids are
eliminated using a suitable suction apparatus. Then the stopped
kinase reaction is applied. After 5 washing steps with 180 .mu.L 1%
trichloroacetic acid in each case the lower half of the dish is
dried (10-20 min at 55.degree. C.) and 25 .mu.L scintillation
cocktail (Microscint, Packard # 6013611) is added. Incorporated
gamma-phosphate is quantified using a Wallac 1450 Microbeta Liquid
Scintillation Counter. Samples without test substance or without
substrate peptide are used as controls. IC.sub.50 values are
obtained using Graph Pad Prism software.
[0156] The anti-proliferative activity of the compounds according
to the invention is determined in the proliferation test on
cultivated human tumour cells and/or in a cell cycle analysis, for
example on NCI--H460 tumour cells. In both test methods the
compounds exhibit good to very good activity, i.e. for example an
EC50 value in the NCI--H460 proliferation test of less than 5
.mu.mol/L, generally less than 1 .mu.mol/L.
[0157] Measurement of the inhibition of proliferation on cultivated
human tumour cells To measure proliferation on cultivated human
tumour cells, cells of lung tumour cell line NCI--H460 (obtained
from American Type Culture Collection (ATCC)) are cultivated in
RPMI 1640 medium (Gibco) and 10% foetal calf serum (Gibco) and
harvested in the log growth phase. Then the NCI--H460 cells are
placed in 96-well flat-bottomed plates (Falcon) at a density of
1000 cells per well in RPMI 1640 medium and incubated overnight in
an incubator (at 37.degree. C. and 5% CO.sub.2). The active
substances are added to the cells in various concentrations
(dissolved in DMSO; DMSO final concentration: 0.1%). After 72 hours
incubation 20 .mu.l AlamarBlue reagent (AccuMed International) is
added to each well, and the cells are incubated for a further 5-7
hours. After incubation the colour change of the AlamarBlue reagent
is determined in a Wallac Microbeta fluorescence spectrophotometer.
EC.sub.50 values are calculated using Standard Levenburg Marquard
algorithms (GraphPadPrizm).
[0158] Cell cycle analyses are carried out for example using FACS
analyses (Fluorescence Activated Cell Sorter) or by Cellomics Array
Scan (CellCycle Analysis).
FACS Analysis
[0159] Propidium iodide (PI) binds stoichiometrically to
double-stranded DNA, and is thus suitable for determining the
proportion of cells in the G1, S, and G2/M phase of the cell cycle
on the basis of the cellular DNA content. Cells in the G0 and G1
phase have a diploid DNA content (2N), whereas cells in the G2 or
mitosis phase have a 4N DNA content.
[0160] For PI staining, for example, 1.75.times.10.sup.6 NCI--H460
cells are seeded onto a 75 cm cell culture flask, and after 24 h
either 0.1% DMSO is added as control or the substance is added in
various concentrations (in 0.1% DMSO). The cells are incubated for
42 h with the substance or with DMSO. Then the cells are detached
with trypsin and centrifuged. The cell pellet is washed with
buffered saline solution (PBS) and the cells are then fixed with
80% ethanol at -20.degree. C. for at least 2 h. After another
washing step with PBS the cells are permeabilised with Triton X-100
(Sigma; 0.25% in PBS) on ice for 5 min, and then incubated with a
solution of propidium iodide (Sigma; 10 .mu.g/ml) and RNAse (Serva;
1 mg/mLl) in the ratio 9:1 for at least 20 min in the dark.
[0161] The DNA measurement is carried out in a Becton Dickinson
FACS Analyzer, with an argon laser (500 mW, emission 488 nm); data
are obtained and evaluated using the DNA Cell Quest Programme
(BD).
Cellomics Array Scan
[0162] NCI--H460 cells are seeded into 96-well flat-bottomed dishes
(Falcon) in RPMI 1640 medium (Gibco) with 10% foetal calf serum
(Gibco) in a density of 2000 cells per well and incubated overnight
in an incubator (at 37.degree. C. and 5% CO.sub.2). The active
substances are added to the cells in various concentrations
(dissolved in DMSO; DMSO final concentration: 0.1%). After 42 h
incubation the medium is suction filtered, the cells are fixed for
10 min with 4% formaldehyde solution and Triton X-100 (1:200 in
PBS) at RT and simultaneously permeabilised, and then washed twice
with a 0.3% BSA solution (Calbiochem). Then the DNA is stained by
the addition of 50 .mu.L/well of 4',6-diamidino-2-phenylindole
(DAPI; Molecular Probes) in a final concentration of 300 nM for 1 h
at RT, in the dark. The preparations are then carefully washed
twice with PBS, the plates are stuck down with black adhesive film
and analysed in the Cellomics ArrayScan using the CellCycle
BioApplication programme and visualised and evaluated using
Spotfire.
[0163] The substances of the present invention are Aurora kinase
inhibitors. On the basis of their biological properties the new
compounds of general formula (I), their isomers and the
physiologically acceptable salts thereof are suitable for treating
diseases characterised by excessive or abnormal cell
proliferation.
[0164] Such diseases include for example: viral infections (e.g.
HIV and Kaposi's sarcoma); inflammatory and autoimmune diseases
(e.g. colitis, arthritis, Alzheimer's disease, glomerulonephritis
and wound healing); bacterial, fungal and/or parasitic infections;
leukaemias, lymphomas and solid tumours (e.g. carcinomas and
sarcomas), skin diseases (e.g. psoriasis); diseases based on
hyperplasia which are characterised by an increase in the number of
cells (e.g. fibroblasts, hepatocytes, bones and bone marrow cells,
cartilage or smooth muscle cells or epithelial cells (e.g.
endometrial hyperplasia)); bone diseases and cardiovascular
diseases (e.g. restenosis and hypertrophy).
[0165] For example, the following cancers may be treated with
compounds according to the invention, without being restricted
thereto: brain tumours such as for example acoustic neurinoma,
astrocytomas such as pilocytic astrocytomas, fibrillary
astrocytoma, protoplasmic astrocytoma, gemistocytary astrocytoma,
anaplastic astrocytoma and glioblastoma, brain lymphomas, brain
metastases, hypophyseal tumour such as prolactinoma, HGH (human
growth hormone) producing tumour and ACTH producing tumour
(adrenocorticotropic hormone), craniopharyngiomas,
medulloblastomas, meningeomas and oligodendrogliomas; nerve tumours
(neoplasms) such as for example tumours of the vegetative nervous
system such as neuroblastoma sympathicum, ganglioneuroma,
paraganglioma (pheochromocytoma, chromaffinoma) and
glomus-caroticum tumour, tumours on the peripheral nervous system
such as amputation neuroma, neurofibroma, neurinoma (neurilemmoma,
Schwannoma) and malignant Schwannoma, as well as tumours of the
central nervous system such as brain and bone marrow tumours;
intestinal cancer such as for example carcinoma of the rectum,
colon, anus, small intestine and duodenum; eyelid tumours such as
basalioma or basal cell carcinoma; pancreatic cancer or carcinoma
of the pancreas; bladder cancer or carcinoma of the bladder; lung
cancer (bronchial carcinoma) such as for example small-cell
bronchial carcinomas (oat cell carcinomas) and non-small cell
bronchial carcinomas such as plate epithelial carcinomas,
adenocarcinomas and large-cell bronchial carcinomas; breast cancer
such as for example mammary carcinoma such as infiltrating ductal
carcinoma, colloid carcinoma, lobular invasive carcinoma, tubular
carcinoma, adenocystic carcinoma and papillary carcinoma;
non-Hodgkin's lymphomas (NHL) such as for example Burkitt's
lymphoma, low-malignancy non-Hodgkin's lymphomas (NHL) and mucosis
fungoides; uterine cancer or endometrial carcinoma or corpus
carcinoma; CUP syndrome (Cancer of Unknown Primary); ovarian cancer
or ovarian carcinoma such as mucinous, endometrial or serous
cancer; gall bladder cancer; bile duct cancer such as for example
Klatskin tumour; testicular cancer such as for example seminomas
and non-seminomas; lymphoma (lymphosarcoma) such as for example
malignant lymphoma, Hodgkin's disease, non-Hodgkin's lymphomas
(NHL) such as chronic lymphatic leukaemia, leukaemic
reticuloendotheliosis, immunocytoma, plasmocytoma (multiple
myeloma), immunoblastoma, Burkitt's lymphoma, T-zone mycosis
fungoides, large-cell anaplastic lymphoblastoma and lymphoblastoma;
laryngeal cancer such as for example tumours of the vocal cords,
supraglottal, glottal and subglottal laryngeal tumours; bone cancer
such as for example osteochondroma, chondroma, chondroblastoma,
chondromyxoid fibroma, osteoma, osteoid osteoma, osteoblastoma,
eosinophilic granuloma, giant cell tumour, chondrosarcoma,
osteosarcoma, Ewing's sarcoma, reticulo-sarcoma, plasmocytoma,
giant cell tumour, fibrous dysplasia, juvenile bone cysts and
aneurysmatic bone cysts; head and neck tumours such as for example
tumours of the lips, tongue, floor of the mouth, oral cavity, gums,
palate, salivary glands, throat, nasal cavity, paranasal sinuses,
larynx and middle ear; liver cancer such as for example liver cell
carcinoma or hepatocellular carcinoma (HCC); leukaemias, such as
for example acute leukaemias such as acute lymphatic/lymphoblastic
leukaemia (ALL), acute myeloid leukaemia (AML); chronic leukaemias
such as chronic lymphatic leukaemia (CLL), chronic myeloid
leukaemia (CML); stomach cancer or gastric carcinoma such as for
example papillary, tubular and mucinous adenocarcinoma, signet ring
cell carcinoma, adenosquamous carcinoma, small-cell carcinoma and
undifferentiated carcinoma; melanomas such as for example
superficially spreading, nodular, lentigo-maligna and
acral-lentiginous melanoma; renal cancer such as for example kidney
cell carcinoma or hypemephroma or Grawitz's tumour; oesophageal
cancer or carcinoma of the oesophagus; penile cancer; prostate
cancer; throat cancer or carcinomas of the pharynx such as for
example nasopharynx carcinomas, oropharynx carcinomas and
hypopharynx carcinomas; retinoblastoma; vaginal cancer or vaginal
carcinoma; plate epithelial carcinomas, adenocarcinomas, in situ
carcinomas, malignant melanomas and sarcomas; thyroid carcinomas
such as for example papillary, follicular and medullary thyroid
carcinoma, as well as anaplastic carcinomas; spinalioma, epidormoid
carcinoma and plate epithelial carcinoma of the skin; thymomas,
cancer of the urethra and cancer of the vulva.
[0166] The new compounds may be used for the prevention, short-term
or long-term treatment of the above-mentioned diseases, optionally
also in combination with radiotherapy or other "state-of-the-art"
compounds, such as e.g. cytostatic or cytotoxic substances, cell
proliferation inhibitors such as for example PLK-inhibitors as
disclosed in WO03/020722 and WO2004/076454, anti-angiogenic
substances, steroids or antibodies.
[0167] The compounds of general formula (1) may be used on their
own or in combination with other active substances according to the
invention, optionally also in combination with other
pharmacologically active substances.
[0168] Chemotherapeutic agents which may be administered in
combination with the compounds according to the invention, include,
without being restricted thereto, hormones, hormone analogues and
antihormones (e.g. tamoxifen, toremifene, raloxifene, fulvestrant,
megestrol acetate, flutamide, nilutamide, bicalutamide,
aminoglutethimide, cyproterone acetate, finasteride, buserelin
acetate, fludrocortinsone, fluoxymesterone, medroxyprogesterone,
octreotide), aromatase inhibitors (e.g. anastrozole, letrozole,
liarozole, vorozole, exemestane, atamestane), LHRH agonists and
antagonists (e.g. goserelin acetate, luprolide), inhibitors of
growth factors (growth factors such as for example platelet derived
growth factor and hepatocyte growth factor, inhibitors are for
example growth factor antibodies, growth factor receptor antibodies
and tyrosinekinase inhibitors, such as for example gefitinib,
imatinib, lapatinib and trastuzumab); antimetabolites (e.g.
antifolates such as methotrexate, raltitrexed, pyrimidine analogues
such as 5-fluorouracil, capecitabin and gemcitabin, purine and
adenosine analogues such as mercaptopurine, thioguanine, cladribine
and pentostatin, cytarabine, fludarabine); antitumour antibiotics
(e.g. anthracyclins such as doxorubicin, daunorubicin, epirubicin
and idarubicin, mitomycin-C, bleomycin, dactinomycin, plicamycin,
streptozocin); platinum derivatives (e.g. cisplatin, oxaliplatin,
carboplatin); alkylation agents (e.g. estramustin, meclorethamine,
melphalan, chlorambucil, busulphan, dacarbazin, cyclophosphamide,
ifosfamide, temozolomide, nitrosoureas such as for example
carmustin and lomustin, thiotepa); antimitotic agents (e.g. Vinca
alkaloids such as for example vinblastine, vindesin, vinorelbin and
vincristine; and taxanes such as paclitaxel, docetaxel);
topoisomerase inhibitors (e.g. epipodophyllotoxins such as for
example etoposide and etopophos, teniposide, amsacrin, topotecan,
irinotecan, mitoxantron) and various chemotherapeutic agents such
as amifostin, anagrelid, clodronat, filgrastin, interferon alpha,
leucovorin, rituximab, procarbazine, levamisole, mesna, mitotane,
pamidronate and porfimer.
[0169] Suitable preparations include for example tablets, capsules,
suppositories, solutions, --particularly solutions for injection
(s.c., i.v., i.m.) and infusion--elixirs, emulsions or dispersible
powders. The content of the pharmaceutically active compound(s)
should be in the range from 0.1 to 90 wt.-%, preferably 0.5 to 50
wt.-% of the composition as a whole, i.e. in amounts which are
sufficient to achieve the dosage range specified below. The doses
specified may, if necessary, be given several times a day.
[0170] Suitable tablets may be obtained, for example, by mixing the
active substance(s) with known excipients, for example inert
diluents such as calcium carbonate, calcium phosphate or lactose,
disintegrants such as corn starch or alginic acid, binders such as
starch or gelatine, lubricants such as magnesium stearate or talc
and/or agents for delaying release, such as carboxymethyl
cellulose, cellulose acetate phthalate, or polyvinyl acetate. The
tablets may also comprise several layers.
[0171] Coated tablets may be prepared accordingly by coating cores
produced analogously to the tablets with substances normally used
for tablet coatings, for example collidone or shellac, gum arabic,
talc, titanium dioxide or sugar. To achieve delayed release or
prevent incompatibilities the core may also consist of a number of
layers. Similarly the tablet coating may consist of a number of
layers to achieve delayed release, possibly using the excipients
mentioned above for the tablets.
[0172] Syrups or elixirs containing the active substances or
combinations thereof according to the invention may additionally
contain a sweetener such as saccharine, cyclamate, glycerol or
sugar and a flavour enhancer, e.g. a flavouring such as vanillin or
orange extract. They may also contain suspension adjuvants or
thickeners such as sodium carboxymethyl cellulose, wetting agents
such as, for example, condensation products of fatty alcohols with
ethylene oxide, or preservatives such as p-hydroxybenzoates.
[0173] Solutions for injection and infusion are prepared in the
usual way, e.g. with the addition of isotonic agents, preservatives
such as p-hydroxybenzoates, or stabilisers such as alkali metal
salts of ethylenediamine tetraacetic acid, optionally using
emulsifiers and/or dispersants, whilst if water is used as the
diluent, for example, organic solvents may optionally be used as
solvating agents or dissolving aids, and transferred into injection
vials or ampoules or infusion bottles.
[0174] Capsules containing one or more active substances or
combinations of active substances may for example be prepared by
mixing the active substances with inert carriers such as lactose or
sorbitol and packing them into gelatine capsules.
[0175] Suitable suppositories may be made for example by mixing
with carriers provided for this purpose, such as neutral fats or
polyethyleneglycol or the derivatives thereof.
[0176] Excipients which may be used include, for example, water,
pharmaceutically acceptable organic solvents such as paraffins
(e.g. petroleum fractions), vegetable oils (e.g. groundnut or
sesame oil), mono- or polyfunctional alcohols (e.g. ethanol or
glycerol), carriers such as e.g. natural mineral powders (e.g.
kaolins, clays, talc, chalk), synthetic mineral powders (e.g.
highly dispersed silicic acid and silicates), sugars (e.g. cane
sugar, lactose and glucose) emulsifiers (e.g. lignin, spent
sulphite liquors, methylcellulose, starch and polyvinylpyrrolidone)
and lubricants (e.g. magnesium stearate, talc, stearic acid and
sodium lauryl sulphate).
[0177] The preparations are administered by the usual methods,
preferably by oral or transdermal route, most preferably by oral
route. For oral administration the tablets may, of course contain,
apart from the abovementioned carriers, additives such as sodium
citrate, calcium carbonate and dicalcium phosphate together with
various additives such as starch, preferably potato starch,
gelatine and the like. Moreover, lubricants such as magnesium
stearate, sodium lauryl sulphate and talc may be used at the same
time for the tabletting process. In the case of aqueous suspensions
the active substances may be combined with various flavour
enhancers or colourings in addition to the excipients mentioned
above.
[0178] For parenteral use, solutions of the active substances with
suitable liquid carriers may be used.
[0179] The dosage for intravenous use is from 1-1000 mg per hour,
preferably between 5 and 500 mg per hour.
[0180] However, it may sometimes be necessary to depart from the
amounts specified, depending on the body weight, the route of
administration, the individual response to the drug, the nature of
its formulation and the time or interval over which the drug is
administered. Thus, in some cases it may be sufficient to use less
than the minimum dose given above, whereas in other cases the upper
limit may have to be exceeded. When administering large amounts it
may be advisable to divide them up into a number of smaller doses
spread over the day.
[0181] The formulation examples which follow illustrate the present
invention without restricting its scope:
Examples of Pharmaceutical Formulations
TABLE-US-00023 [0182] A) Tablets per tablet active substance
according to formula (1) 100 mg lactose 140 mg corn starch 240 mg
polyvinylpyrrolidone 15 mg magnesium stearate 5 mg 500 mg
[0183] The finely ground active substance, lactose and some of the
corn starch are mixed together. The mixture is screened, then
moistened with a solution of polyvinylpyrrolidone in water,
kneaded, wet-granulated and dried. The granules, the remaining corn
starch and the magnesium stearate are screened and mixed together.
The mixture is compressed to produce tablets of suitable shape and
size.
TABLE-US-00024 B) Tablets per tablet active substance according to
formula (1) 80 mg lactose 55 mg corn starch 190 mg microcrystalline
cellulose 35 mg polyvinylpyrrolidone 15 mg sodium-carboxymethyl
starch 23 mg magnesium stearate 2 mg 400 mg
[0184] The finely ground active substance, some of the corn starch,
lactose, microcrystalline cellulose and polyvinylpyrrolidone are
mixed together, the mixture is screened and worked with the
remaining corn starch and water to form a granulate which is dried
and screened. The sodiumcarboxymethyl starch and the magnesium
stearate are added and mixed in and the mixture is compressed to
form tablets of a suitable size.
TABLE-US-00025 C) Ampoule solution active substance according to
formula (1) 50 mg sodium chloride 50 mg water for inj. 5 ml
[0185] The active substance is dissolved in water at its own pH or
optionally at pH 5.5 to 6.5 and sodium chloride is added to make it
isotonic. The solution obtained is filtered free from pyrogens and
the filtrate is transferred under aseptic conditions into ampoules
which are then sterilised and sealed by fusion. The ampoules
contain 5 mg, 25 mg and 50 mg of active substance.
* * * * *